Eagle s minimum essential medium

Unless otherwise indicated, all chemicals used for preparation of the scaffold and cell sheet decellularization were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dichloromethane (DCM) was purchased from VWR Chemicals (Radnor, PA, USA).
All cells used were purchased from ATCC, except that the mesenchymal stem cells (MSC) were purchased from Innoprot (Bizkaia, Spain), and the human chondrocyte cells were purchased from Cell Application INC (San Diego, CA, USA). Culture reagents, including fetal bovine serum (FBS), penicillin–streptomycin, and trypsin, were purchased from Life Technologies (Carlsbad, CA, USA). The culture media were purchased from the following sources: rat mesenchymal basal medium and its growth supplement, Cell Application Inc.; Eagle’s minimum essential medium, ATCC; Dulbecco’s modified Eagle medium/nutrient mixture F-12, Gibco (Billings, MT, USA); Dulbecco’s modified Eagle medium, Gibco.
The analytical reagents were purchased from the following sources: CellTiter Blue cell viability assay, Promega (Madison, WI, USA); Quant-iT™ PicoGreen^® dsDNA, Invitrogen (Waltham, MA, USA); BCA assay kit, Thermo Fisher Scientific (Waltham, MA, USA).

We obtained human prostate cancer PC3 (CRL‐1435), DU‐145 (HTB‐81), and LNCaP (CRL‐1740) cells from ATCC (Rockville, MD, USA). The PC3 cells were incubated in F‐12 K, 1X (Ham's F‐12 K Nutrient Mixture, Kaighn's Mod.) with L‐glutamine (10‐025‐CV) purchased from Corning Incorporated (Oneonta, NY, USA). The DU‐145 cells were incubated in Eagle's Minimum Essential Medium (30–2003) purchased from ATCC. The LNCaP cells were grown in RPMI‐1640 (30–2001) media purchased from ATCC. For each cell line, we added 10% fetal bovine serum (Corning, 35‐010‐CV) and 1% antibiotic (Hyclone, SV30010) and kept them in a 37°C humidified 5% CO2 incubator from passages 4–30. Docetaxel‐Resistant Clones were previously prepared in the lab as reported by Asay 2020.⁶

Murine melanoma cells (B16F10) and human glioblastoma cells (U87MG) as well as Dulbecco’s Modified Eagle’s Medium (DMEM) and Eagle’s Minimum Essential Medium (EMEM) were purchased from ATCC (Wesel, Germany). Fetal calf serum (FCS) was obtained from Bio&SELL (Feucht, Germany); phosphate-buffered saline (PBS), 1,10-phenanthroline, tris(hydroxymethyl)aminomethane hydrochloride (Tris·HCl) and manganese chloride (MnCl₂) were obtained from Sigma-Aldrich (Taufkirchen, Germany); penicillin/streptomycin (pen/strep) and 0.25% Trypsin with 0.02% EDTA-solution in PBS were obtained from Gibco (Schwerte, Germany); 2-(4-(2-hydroxyethyl)-1-piperazinyl)-ethanesulfonic acid (HEPES) was obtained from Gerbu (Heidelberg, Germany); bovine serum albumin (BSA), sodium chloride (NaCl), calcium chloride (CaCl₂) and magnesium chloride (MgCl₂) were obtained from Carl Roth (Karlsruhe, Germany). [¹²⁵I]I-NDP (NEX352, 81.4 GBq/μmol) and [¹²⁵I]I-echistatin (NEX083, 81.4 GBq/μmol) were purchased from PerkinElmer (Rodgau, Germany). γ-counting was performed using a 2480 Wizard² gamma counter system from PerkinElmer.

Rat glioma (C6) was cultured in Ham’s F12 medium supplemented with 15% horse serum and 2.5% fetal bovine serum. Human Caco-2 was cultured in Eagle’s minimum essential medium with 20% fetal bovine serum. Human neuroblastoma (SH-SY5Y) was grown in Dulbecco’s modified Eagle medium with 10% fetal bovine serum and 1% 200 mM l-glutamine. All cells were maintained in 5% CO₂ air atmosphere, 37°C. Upon confluence in flasks, cells were subcultured on glass cover slips in 24-well cell culture dishes. For Cell IQ and gene array observations, cells were grown on well bottoms without cover slips. C6, Caco-2, SH-SY5Y, Eagle’s minimum essential medium, Ham’s F12 medium, horse serum, and fetal bovine serum are from ATCC (Manassas, VA, USA). fetal bovine serum, Dulbecco’s modified Eagle media, trypsin–EDTA, and l-glutamine are from Invitrogen (Stockholm, Sweden). T-75 cm² cell culture flasks and polystyrene 24-well cell culture dishes are from Nunc (Roskilde, Denmark). Glass cover slips (∅ 13 mm, thickness = 0.15 mm) are from BergmanLabora (Danderyd, Sweden). All controls/sham cultures were treated likewise but not exposed to in vitro trauma.

Formic acid was purchased from Sigma Aldrich (Steinheim, Germany), while acetonitrile (LC-MS grade) and methanol (HPLC grade) were acquired from Merck (Darmstadt, Germany). A Milli-Q Simplicity 185 water filtration system (Millipore, Milford, MA, United States) generated ultrapure water. Regarding the antioxidant activity evaluation, lipoxidase from soybean (type I-B), linoleic acid, quercetin, and dimethyl sulfoxide (DMSO) were purchased from Sigma Aldrich (Steinheim, Germany), and acetate buffer 0.1 M (pH 5.25) was prepared by combining sodium acetate with acetic acid (Sigma Aldrich) until the required pH was reached. Similar outcomes were achieved by combining boric acid (Sigma-Aldrich) and NaOH (1 N), thus obtaining borate buffer (pH 9). In addition, the ferrous sulfate solution in 0.2 M hydrochloric acid and the 5 mM ferrozine solution were prepared using chemicals purchased from Sigma Aldrich (Steinheim, Germany).
Considering the cytotoxicity assays, Dulbecco’s Modified Eagle’s Medium, as well as the penicillin/streptomycin mixture, were acquired from Sigma Aldrich, while Eagle’s Minimum Essential Medium was purchased from ATCC. Moreover, the fetal bovine serum was acquired from ThermoFisher Scientific, Boston, MA, USA.